3-O-C12-HSL阻碍Mo-DCs成熟过程中lncRNA表达谱的变化

Differential expression profile of long non-coding RNA in 3-O-C12-HSL-inhibted Mo-DCs maturation

摘要:

目的：观察铜绿假单胞菌（Pseudomonas aerltginosa，PA）分泌的信号分子N-3-氧代十二烷酰-L-同型丝氨酸内酯（N-3-oxododecanoy1-L-homoserine lactone，3-O-C12-HSL）阻碍单核细胞来源的树突细胞（monocytes derived-dendritic cells，Mo-DCs）成熟过程中长链非编码RNA（long non-coding RNA，lncRNA）表达谱的变化情况。方法：采用免疫磁珠法分选人外周血CD14+单核细胞，经重组人粒-单核细胞集落刺激因子和重组人白细胞介素-4诱导分化为Mo-DCs。不同因素处理Mo-DCs，实验组加入终浓度0.1 μg/mL的LPS和终浓度为40 μmol/L的3-O-C12-HSL，对照组加入相应体积的含0.1%DMSO的PBS，模型组加入终浓度0.1 μg/mL的LPS。利用lncRNA芯片技术检测对照组、模型组和实验组间lncRNA表达谱的差异。对筛选数据进行分析处理，利用散点图分析2倍以上差异lncRNA在各处理因素间的总体变异情况，利用聚类分析研究表达差异5倍以上lncRNA的聚类情况。结果：与对照组和模型组相比较，实验组筛选出2倍以上表达差异的lncRNA为1 386条，其中上调表达的548条，下调表达的838条。筛选出具有5倍以上表达差异的lncRNA共153条，占2倍表达差异lncRNA的11.04％，其中上调表达的22条，下调表达的131条。散点图发现2倍表达差异的lncRNA总体分布呈集中趋势，存在特征性改变。聚类分析发现5倍表达差异的lncRNA能根据处理因素进行有效的分层聚类。结合GO分析和mRNA通路分析，差异lncRNA的相关基因可富集至PPAR信号通路、钙信号通路和NF-κB信号通路中。结论：3-O-C12-HSL作用于Mo-DCs后，lncRNA表达谱发生了特征性变化，表达差异的lncRNA可能参与了3-O-C12-HSL阻碍Mo-DCs成熟过程。

Abstract:

Objective：To analyze the expression profile variation of long non-coding RNA（lncRNA） in 3-O-C12-HSL-inhibted Mo-DCs maturation. Methods：Human CD14+ monocytes were separated by immunomagnetic beads and induced by hIL-4 and hGM-CSF. Experimental group was stimulated with 40 μmol/L 3-O-C12-HSL and 0.1 μg/mL LPS；control group was stimulated with 0.1% DMSO and model group was stimulated with 0.1 μg/mL LPS. lncRNA chip was used to analyze the differential expression among control group，model group and experimental group. Significant differe-ntial lncRNA（P<0.05） was analyzed and scatter plot was used to study the variance in 2 times differential lncRNA. Clustering analysis was used to study the 5 times differential lncRNA. Results：Comparing with those of control group and model group，1 386 lncRNA which have more than 2 times variation and significant differences by statistical analysis were selected out，including 548 up-regulated lncRNA and 838 down-regulated lncRNA. Meatime，153 lncRNA accounting for 11.04％ of all differential lncRNA had more than 5 times variation in differ－ence groups. Characteristic change were displayed among 2 times differential lncRNA in scatter plot analysis. The hierarchical clustering results displayed that 5 times lncRNA clustering efficient in different groups. According to GO analysis and mRNA pathway analysis，PPAR signaling pathway，calcium signaling pathway and NF-κB signaling pathway were enriched with differential lncRNA associated genes. Conclusion：lncRNA displaying characteristic change may involve in 3-O-C12-HSL-inhibted Mo-DCs maturation.